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Master's Dissertation
DOI
10.11606/D.46.2005.tde-23112015-112115
Document
Author
Full name
Flavia Garcia
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2005
Supervisor
Committee
Catalani, Luiz Henrique (President)
Medeiros, Marisa Helena Gennari de
Russo, Momtchilo
Title in Portuguese
Formação de oxigênio singlete O2 (1Δg) por fagócitos
Keywords in Portuguese
Antraceno derivado
Fagócitos mononucleares (Estudo)
Oxigênio (Formação)
Oxigênio molecular singlete
Radicais livres
Sistemas biológicos
Abstract in Portuguese
Neste trabalho avaliamos a formação de oxigênio singlete in vitro em fagócitos, (células mononucleares e neutrófilos) isolados de sangue periférico humano, e eosinófilos, de lavado bronco alveolar de camundongos balb/c, ativados por estímulo partículado: zimosan opsonizado contendo o 9,10difenilantraceno (DPA) adsorvido como sonda captadora de 1O2. Por este método, a formação do 1O2 pode ser verificada pela formação do 9,10-difenilantraceno endoperóxido (DPAO2), que é detectado por HPLC. Observamos, que os fagócitos formam 1O2 e que esta formação parece ocorrer de forma diferenciada para os dois tipos celulares (neutrófilos e células mononucleares). Visando ampliar os estudos anteriores sobre o papel da melatonina (MLT) no processo inflamatório, foi testado seu efeito em fagócitos e a relação na produção de 1O2 destas células. Observamos que MLT inibe a formação de 1O2 totalmente no caso de neutrófilos e parcialmente no caso de células mononucleares e eosinófilos. Paralelamente, foi desenvolvida a síntese de um novo captador químico de 1O2, o éster 9,10-antracenil-3-bispropionato de etila (ABPE), cuja finalidade principal é o acúmulo no interior da célula, depois de sofrer hidrólise enzimática. Esta sonda, terá facil acesso ao interior das células em sua forma ester. Este novo captador de 1O2 foi testado em células mononucleares e neutrófilos estimulados de formas diferentes: via receptor independente e dependente. Os resultados demonstraram produção equivalente de 1O2 nestes fagócitos.
Title in English
Singlet oxygen formation O2 (1Δg) by phagocytes
Keywords in English
Anthracene derivative
Biological systems
Free radicals
Mononuclear phagocytes (Study)
Oxygen (Formation)
Singlet molecular oxygen
Abstract in English
In this study, we evaluated the singlet oxygen (1O2) formation in vitro from phagocytes (neutrophils and mononuclear cells) isolated from human blood cells and eosinophils isolated from bronchoalveolar lavage fluid of mice balb/c activated, by opsonized zymosan. To determine whether singlet oxygen is produced by phagocytes, zymosan particles were coated with a specific chemical trap for 1O2, 9,10-diphenylanthracene (DPA). The production of 1O2 was followed using HPLC, to measure its product, 9,10-diphenylanthracene endoperoxide (DPAO2). We also noticed that the 1O2 production occurs at different levels of for two cell types, neutrophils and mononuclear cells. In order to broaden previous studies on the role of melatonin (MLT) in inflammatory processes, its effect was tested in phagocytes was tested in relation to 1O2 formation by these cells. We observed that MLT inhibits the 1O2 formation totallymt neutrophils and partiallym mononuclear cells and eosinophils. At the some time, it was also developed the synthesis of a new probe for 1O2, the 9,10-anthracene-bis-3-ethyl-propionate (ABEP), with the purpose to accumulate inside the cells, after its enzymatic hydrolysis. This probe presents easy acess to the inferior of the cells in its ester form. This new probe for trapping 1O2 was tested in mononuclear cells and neutrophils stimulated in two ways: via independent and dependent receptor. The results showed equivalent production of 1O2 for both cell types.
 
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Publishing Date
2015-11-23
 
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